PromoCell’s crypreserved human monocyte-derived macrophages (hMDM) are convenient and easy-to-handle. The thawed cells plate into all tissue culture vessel formats and can be maintained as adherent, biologically functional cultures for several weeks. Optionally, user-customizable activation of the cells can be performed.

Cryopreserved human macrophages are produced from monocytes in PromoCell’s well-proven M2 Macrophage Generation Media DXF and are available as non-activated, fully qualified M2- (M-CSF) polarized cells. After 9-10 days of differentiation, the M2 macrophages are cryopreserved using PromoCells’s proprietary, defined and animal-free freezing medium, Cryo-SFM. Each cryo vial contains more than 1.5 million or 5 million viable cells after thawing, respectively.

For the M2 Macrophage Generation Medium, it is extremely important that the shelf life of 2 weeks (after addition of the supplements) is not exceeded; the yield will quickly decrease thereafter. It is best to use the M2 medium as fresh as possible to avoid discrepancies between M1 and M2 yield.

a) We usually perform macrophage differentiation in T75 flasks and 6-well plates. We haven’t tested differentiation in smaller formats. But we assume it will be problematic to thoroughly wash the surface of the wells to remove non-adherent cells after the attachment phase. b) Detachment of the mature macrophages is possible but re-attachment can lead to significant cell loss (30-50%).
Please also keep in mind that working in 96/384 well Format has some inherent drawbacks (e.g. Evaporation of media, dry wells, etc).

It is not recommended to leave the blood cells in the Monocyte Attachment Medium for longer than 1.5-2 hrs. The medium was developed for (short-term) attachment of the monocytes and does not provide nutrients for a longer time period. Leaving the cells in Monocyte Attachment Medium for a longer time or even overnight will induce apoptosis and lead to the loss of the cells.
If necessary, you can reduce the incubation time to 1 hr. In this case, it is advisable to equilibrate the media in the incubator before so that you can immediately and directly add the appropriate amount of PBMC suspension.

The Macrophage Detachment Solution (C-41330) directly affects the cell membrane. HSA in the Wash Buffer supports Regeneration of the cell membrane and protects the cells during the critical phase directly after detachment from detrimental effects.

Spinning the cells for 15 min at 350 x g has been proven and tested by PromoCell Research & Development. The QC department uses these settings during testing. Any lower centrifugation value (g-force and/or time) will lead to significant cell loss by means of non-sedimented, but intact, macrophages.

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It is not necessary to use coated flasks for (most of) our Normal Human Cells but it can be done. As coating with extracellular matrix proteins can affect cellular metabolism, it is recommended to use the same coating material for a complete set of experiments.
Cells that need to be grown on coated dishes:

Usually, we do not recommend specific plasticware but we have found that with PromoCell’s Dendritic Cell and Macrophage Generation Media, choice of plasticware can have a lot of an influence. For our Macrophage Generation Media DXF we recommend the Nunc plasticware with Nunclon surface – as not only will the detachment efficiency vary (up to 20%), but also the efficiency of the differentiation process itself may be altered.

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No, our Growth/Differentiation/Generation Media are supplied as ready-to-use media. After addition of the provided supplements to the basal medium, the media are complete and no further supplementation with serum or other growth factors is required.Exceptions:

Usually, we do not recommend specific plasticware but we have found that with PromoCell’s Dendritic Cell and Macrophage Generation Media, choice of plasticware can have a lot of an influence. For our Dendritic Cell Generation Media we recommend to use tissue culture vessels from BD FalconTM.

Many rules in biology exist, and even more exceptions to these rules. For years, immunologists have tried to classify macrophages into two subtypes, classically activated M1, and alternatively activated M2 macrophages. However, with the growing number of activated macrophage subtypes needing description, established nomenclature falls short. Researchers are seeking an alternative to the traditional black-and-white principle….

One cell type can fight disease, help form blood vessels, regulate cell stability— and more. Macrophages are true masters of multitasking. In our body they are the first line of defense against invading bacteria, fungi and viruses. But they are not only a key element of our immune defense; they can also play a role in destroying tissue,…